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BACKGROUND: Chemokine (C-C motif) receptor 6 (CCR6) is present in sperm and plays a significant role in sperm motility and chemotaxis acting in the reproductive tracts. However, the expression and functional significance of CCR6 in testis are still poorly understood, especially in the process of spermatogenesis. METHODS AND RESULTS: CCR6 was expressed in spermatogenic cell lines and its expression was shown in an age-dependent upregulation manner from puberty to adulthood in mouse testis. Immunostaining results confirmed the localization of CCR 6 in testis. Further chemotaxis assays demonstrated that spermatogenic cells GC-1 and -2 exhibited a directional movement toward CCR6-specific ligand such as CCL20 or Sertoli cells in vitro. CONCLUSIONS: The present findings indicate that CCR6 is involved in the chemotaxis of spermatogenic cells in vitro and promotes chemotaxis under non-inflammatory conditions during normal spermatogenesis.

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PURPOSE: We compared and contrasted the structure of the gubernaculum testis in fetuses with prune belly syndrome and normal controls. MATERIALS AND METHODS: We studied a total of 6 gubernacula from 3 male fetuses with prune belly syndrome and a total of 14 from 7 male fetuses without an anomaly. Gubernacular specimens were cut into 5 µm sections and stained with Masson trichrome to quantify connective tissue and smooth muscle cells, with Weigert stain to observe elastic fibers and with picrosirius red with polarization to observe collagen. Immunohistochemical analysis was done with tubulin to observe the nerves. Images were captured with a BX51 microscope and DP70 camera (Olympus®). Stereological analysis was done with Image-Pro and ImageJ (MediaCybernetics®) using a grid to determine volumetric density. Means were statistically compared with the Mann-Whitney test. All tests were 2-sided with p <0.05 considered statistically significant. RESULTS: Prune belly syndrome fetuses were at 17 to 31 weeks of gestation and control fetuses were at 12 to 35 weeks of gestation. Quantitative analysis showed no difference in the volumetric density of smooth muscle cells in prune belly syndrome vs control gubernacula (mean 15.70% vs 19%, p = 0.2321). Collagen fiber analysis revealed a predominance of green areas in prune belly syndrome gubernacula, suggesting collagen type III, and a predominance of red areas in control gubernacula, suggesting collagen type I. Elastic fibers were significantly smaller in prune belly syndrome gubernacula than in control gubernacula (mean 14.06% vs 24.6%, p = 0.0190). Quantitative analysis demonstrated no difference in the volumetric density of nerves in prune belly syndrome or control gubernacula (mean 5.200% vs 3.158%, p = 0.2302). CONCLUSIONS: The gubernaculum in fetuses with prune belly syndrome had altered concentrations of collagen and elastic fibers. These structural alterations could be one of the factors involved in cryptorchidism in prune belly syndrome.

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REASONS FOR PERFORMING STUDY: Cryptorchidism affects 2-8% of male horses and the affected testis undergoes a disruption of normal spermatogenesis. The underlying molecular changes are poorly understood in the cryptorchid equine testis. OBJECTIVES: Compare the expression of anti-Müllerian hormone (AMH), anti-Müllerian hormone receptor (AMHR2), androgen receptor (AR), cyclin kinase inhibitor (CDKN1B), connexin 43 (Cx43), 3ß hydroxysteroid dehydrogenase/Δ(5) -Δ(4) - isomerase (3ßHSD), P450c17 hydroxylase/lyase (P450c17) and cytochrome P450 aromatase (P450arom) in the undescended testis of cryptorchid stallions with that of normal stallions. METHODS: Undescended, abdominal testes from four cryptorchid stallions between 2 and 3 years of age were collected during routine castrations along with normally descended testes from normal stallions between 2 and 3 years of age (n = 7). Samples were analysed by immunohistochemistry and quantitative real-time PCR. RESULTS: Cryptorchid testes had increased AMH and AMHR2 immunolabelling when compared with normal testes, which indicates failure of maturation of Sertoli cells and/or lack of testosterone suppression. Failure of Sertoli cell maturation in the cryptorchid testis may also be attributed to AR abnormalities and/or a consequence of lack of testosterone suppression due to decreased 3ßHSD. Cyclin-dependent kinase (CDKN1B) was not expressed in Sertoli cells of cryptorchid testes suggesting that Sertoli cells are still proliferating, which is also a characteristic of the immature testis. In addition, Cx43 expression is decreased in the cryptorchid testis, indicating a disruption in intercellular communication. CONCLUSIONS: Undescended testes of cryptorchid horses present characteristics of immaturity suggesting that the failure of Sertoli cell maturation may be a consequence of cryptorchidism. POTENTIAL RELEVANCE: This study provides a better understanding of the effect of cryptorchidism on testicular function in stallions.

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PURPOSE: We evaluated extracellular matrix remodeling in human fetal and cryptorchidic gubernacula. METHODS: Gubernacula were obtained from 40 normal human fetuses aged 15-29 weeks postconception (WPC) and from 39 children aged 1.3-10 years who had been submitted to orchiopexy. Total collagen and glycosaminoglycan (GAG) concentrations were assessed as µg hydroxyproline and µg hexuronic acid per mg dry gubernacular tissue, respectively, and proportions of sulfated GAG were determined by agarose gel electrophoresis. RESULTS: Fetal age correlated with collagen (r = 0.77, P < 0.001) and GAG (r = -0.83, P < 0.01) concentrations, which varied from 10 to 50 µg/mg and 7.1-2.5 µg/mg, respectively. Collagen and GAG concentrations in cryptorchidic gubernacula varied from 80.0 to 120.0 µg/mg and from 0.8 to 1.0 µg/mg, respectively. These values did not correlate with patient's age, but when the testis was located more proximally, collagen content was lower. At 15-18 WPC, GAG consisted of 57.3% ± 13.5% chondroitin sulfate and 28.2% ± 10.1% dermatan sulfate, and at 25-28 WPC, 42.7% ± 8.7% and 71.8% ± 11.3%, respectively. GAG in postnatal gubernacula consisted mostly of dermatan sulfate. CONCLUSIONS: From the 15th to the 29th WPC, the extracellular matrix of the gubernaculum undergoes extensive remodeling and this may contribute to testicular descent. Cryptorchidic gubernacula are markedly more fibrous than the corresponding fetal tissue, change little after birth, and have a lower collagen concentration when the undescended testis is abdominal in position.

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The aim of the present study was to examine the effects of experimental cryptorchidism on rat testicular phospholipids and neutral lipids that contain long-chain (C(18)-C(22)) and very long-chain (VLC) (C(24)-C(32)) polyunsaturated fatty acids (PUFA). The weight of the cryptorchid testis was nearly half that of the contralateral control at postsurgical Days 7-10 owing to the depletion of germ cells. Concomitantly, the amounts of major glycerophospholipids (GPL) and sphingomyelin (SM) per testis decreased. Both these lipids lost their characteristic long-chain and very long-chain PUFA, notably 22:5n-6 and 28:4n-6, respectively, which suggests that these species are linked to the membranes of germ cells. In contrast, the amounts and concentrations of triglycerides (TG; triacylglycerols and 1-O-alkyl-2,3-diacylglycerols) and cholesterol esters (CE) increased several fold in the surviving cells (mainly Sertoli cells) in the cryptorchid testis. All these neutral lipids, but especially CE, accumulated large amounts of the major PUFA of the testis, 22:5n-6, as well as pentaenes with longer carbon chains (i.e., 24:5n-6 in TG and 28:5n-6 in CE). This accretion suggests that neutral lipids may store preformed PUFA coming from dying germ cell GPL and also VLCPUFA no longer needed as a source of PUFA destined to assemble new germ cell GPL. The lipid adjustments observed in cryptorchidism suggest a possible role for Sertoli cell CE in the turnover and conservation of PUFA within seminiferous tubules.

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